Start-up
1. Turn MASTER switch on Belkin Surge-
Master. This will turn on the LUDL stage
controller, microscope stand and the
Olympus Microfire color CCD camera.
2. Turn ON X-Cite 120 Hg lamp if needed.
Note: The X-Cite 120 must stay ON a
minimum of 20 minutes. It also requires
20 minutes to cool down. If you turn it
off, the internal logic chip will not let the
system arc until it is cooled down.
3. Log in to the computer.
4. Load the Stereo Investigator or Neu-
rolucida software (shortcut on desktop
after training).
Once the software is up:
Set reference point in Live image in
software. This will make the live window
and mouse curser more responsive to
movement.
Load a sample and find
focus...
1. Load a sample: Rotate the 5x lens into
position, then press the stage speed
button on the joystick to move the stage
out. Load your sample. Move the sample
back under the objective. Push in the
push/pull rod to deflect light to the
binocular tube. Use coarse focus on the
right side of the stand and/or the fine
focus control on the joystick controller.
See joystick image to right for help.
2. Select objective in the software so
the software knows what objective you
are using.
3. Select Imaging Method:
For conventional brightfield viewing:
• Rotate the Condenser Turret Disk to the
H position.
• Swing out the polarizing filter located
under the condenser and remove the
full wave lambda plate if present.
• Align the microscope for Koehler Il-
lumination.
For Transmitted dark field contrast
microscopy:
Using the 10x, 5x or 2.5x objective, you
can set the Condenser Turret Disk to D
after doing the steps above including
Koehler Illumination.
For widefield fluorescence viewing:
• Press the RL Key.
• Set Reflector Turret to (2) Eyes DUAL
470EX & 560EX/515EM & 620EM
• Adjust the Incident Iris Stop Slider until
light intensity is adequate.
• Find Focus preferably on a dye that
emits in the green range (500-530nm).
• Pull out the Push/Pull rod to deflect
light to the camera.
Start-up
1 . . . . . . . . . Turn ON Master Switch
2 . . . . . . . . . . Log on to Computer
3 . . . . . . . . . . . . . Load Software
Optional . . . . Turn ON X-Cite Hg Lamp
Align Optical Components
1 . . . . . . . . . . Koehler Illumination
Calibrate System
1 . . . . . . . . . . . . . Grid Tune Lens
2 . . . . . Parcentric Parfocal Calibration
Run Experiment
1 . . . . . . . . . . . . . Set up Subject
2 . . . . . . . . . .Set scope to Low Mag
3 . . . . . . . .Trace Region(s) of interest
4 . . . . . . . . . Set scope to High Mag
5 . . . . . . . Define Counting Frame Size
6 . . . . . . . . . Define SRS Grid Layout
7 . . . . . . . . .Define Disector Options
8 . . . . . . . . Save Sampling Paramters
9 . . . . . . . Select Markers for Counting
10 . . . . . . . . . . . . Count Objects
11 . . . . . . . . View Sampling Results
Fluorescent Imaging
Turn ON X-Cite 120 UV lamp
Note: For Fluorescence imaging:
Check the Reflector turret is set to desired
excitation/emission.
•
Reflector Turret set to:
(1) Transmitted Light/Transmitted Pol.
(2) Eyes DUAL 470EX & 560EX/515EM & 620EM
(3) Triple Camera only Triple EX & EM
(4) Quad Camera only single BP
(5) Blank
(6) Reflected light
•
Set Reflected light shutter to ON
and transmitted light shutter to OFF
Transmitted Light Imaging
The green LED located on the lower right
side of the microscope base indicates both
microscope power and that the transmitted
light shutter is ON. The light intensity control
knob will attenuate halogen power.
Set the Reflector Turret to position (1) Trans-
mitted Light/Transmitted Polarized light
Set the condenser to H for Bright-field viewing
or D for Dark-field. Remember, depending on
the objective chosen, you may have to swing
out or swing in the condenser front lens. 2.5x
and 5x the objective front lens should be
swung out.
For polarized light imaging a full wave com-
pensator is available.
